[Preparation of anti-ASNS monoclonal antibody and detection of ASNS expression in tumor].

Abstract

AIM:

To prepare and characterize monoclonal antibodies against ASNS for the research of ASNS function.

METHODS:

To induce the expression of the fusion protein MS2-ASNS, Hybridomas were generated by the fusion with Sp2/0 myelomas and spleen cells, which were obtained from mice immunized with MS2-ASNS recombinant proteins. The specificity and titer of mAb were identified by ELISA methods, and then used to detect the affinity of ASNS in cancer cells by Western blot. The expression of ASNS was detected in some cancer lines and tissues by IHC.

RESULTS:

Two hybridoma cell lines F4-15 and F4-16, which stably secret anti-ASNS mAbs were produced. Both cell lines produce IgG2a monoclonal antibody. ELISA demonstrated that anti-ASNS mAbs had high specificity and titer. The titers of anti-ASNS mAbs produced by hybridoma cell lines were up to 1:5×10(5);. Western blot demonstrated that ASNS was expressed in some cancer lines, including human lymphoma cell line K562 and cervical cancer cell line HeLa. The expression of ASNS was also detected by IHC in several tumor cell lines, such as stomach cancer cell line SGC-7901 and liver cancer cell lines SMMC-7721, BEL-7402, HepG2 as well as lung and esophageal carcinoma tissue.

CONCLUSION:

The monoclonal antibodies against ASNS have been successfully prepared, which provides a tool for the following research of nasal type NK/T cell lymphoma.

• Authors: Wei Shi, Qing-yun Zhang, Zhan-sheng Jia, Jian-jun Xu, Ya-ming Wang, Wei-ben Yong
• Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi) Vol. 27 Issue 1 Pg. 74-7 (Jan 2011) ISSN: 1007-8738 [Print] China
• PMID: 21208570 (Publication Type: English Abstract, Journal Article)

Chemical References

• Antibodies, Monoclonal
• Antigens, CD
• Immunoglobulin G
• Membrane Proteins
• Recombinant Fusion Proteins
• ADAM Proteins
• Adam8 protein, mouse
• Aspartate-Ammonia Ligase

Topics

• ADAM Proteins (genetics, immunology)
• Animals
• Antibodies, Monoclonal (biosynthesis, immunology, isolation & purification)
• Antibody Specificity
• Antigens, CD (genetics, immunology)
• Aspartate-Ammonia Ligase (biosynthesis, genetics, immunology)
• Blotting, Western (methods)
• Cell Line, Tumor
• Enzyme-Linked Immunosorbent Assay (methods)
• Female
• HeLa Cells
• Humans
• Hybridomas (immunology)
• Immunoglobulin G (biosynthesis, immunology, isolation & purification)
• K562 Cells
• Membrane Proteins (genetics, immunology)
• Mice
• Mice, Inbred BALB C
• Neoplasms (chemistry, enzymology)
• Recombinant Fusion Proteins (biosynthesis, genetics, immunology)